| Objective: To investigate the regulation of the probiotics on tightjunction protein Claudin-1of intestinal mucosal in rats with trinitro-benzen-sulfonic acid (TNBS) induced colitis and further exploration of the effect ofprobiotics protecting on intestinal mucosal barrier with colitis in experimentalrats.Methods: Twenty-eight male of SD rats were divided3groups, controlgroup (A group, n=8), without Probiotics treatment group (B group, n=10),Probiotics treatment group (C group, n=10). To establish the model ofTNBS-induced rat colitis: The SD rats fasting24hours before operation,Witha diameter of about2mm and length about12cm silicone tube gently insert adepth of about8cm into anus of group B and group C with containing0.85mlsolution with80mg/kg TNBS equal volume of50%ethanol solution slowlyinto the colon, and let rats keep anus raise at30degree, The Control groupenema with an equal volume of saline into the colon, Each rat of the controlgroup and the group B daily with1ml distilled water enema once everyday.Probiotics group, each rat enema from1stto7thday with Golden Bifid2Copowder with distilled water preparation1ml enema. Rats in each group withnormal feeding. Observation eating, activity, stool and weight of ratseveryday. The8thday, after sodium pentobarbital anesthesia and take theinner canthus blood to determination plasma D-lactic acid TNF-alpha andserum IL-8. Take the same part of2cm colon tissue of each rat, formalin fixed for HE staining and immunohistochemical assay of Claudin-1protein expression;The remaining specimens were pre-fixed with3%glutaraldehyde for observation the ultrastructural changes colon intestinal tissue with transmissionelectron microscopy (TEM).Results: General condition: After modeling the first day, group B ratssuffered from diarrhea with gross bloody stool or fecal occult blood testpositive or strong positive, rat mental activity badly, eating less, weight loss,the second day to5thday diarrhea most severity, the5thto6thday after theblood stool will gradually reduce, eating recovery. The group C rats aftermodeling the first day had loose stool or unshapen, mental activity somewhatless, eating less, stool occult blood positive. The5thday, the diarrhea, occultblood was reduced, activity increased, eating recovery. The group A rats aftermodeling the first day mental activity and diet back to normal. Diseaseactivity index (DAI)：group A、group B and group C DAI were0.49±0.43、3.17±0.43and2.08±0.57, the difference between groups were statisticallysignificant (P <0.05); Histologic injury score: Group A、group B and group Cwere1.00±0.76、3.1±1.20and2.1±1.1，the difference between groupswere statistically significant (P<0.05)；TNF-a level: group A、B and C groupswere35.13±18.37、172.72±17.07and57.20±19.66, the difference betweengroups were statistically significant (P<0.05); Plasma D-lactate levels: groupA、group B and group C respectively5.267±7.38、12.66±2.68and11.50±7.31，group B and group C compared with group A differences werestatistically significant (P<0.05); but group B compared with group Cdifference was not significant (P>0.05). Serum IL-8levels: group A、B and Cgroups were3.32±2.28、9.18±2.04and5.84±2.37, the difference between groups were statistically significant (P<0.05); Colon intestinal epithelial celltight junction protein Claudin-1protein expression: Group A、groupB andgroup C immunohistochemistry results showed that the differences werestatistically significant between the groups (P<0.05). TEM results showedthat intestinal villus of group A were intensive, arranged in neat rows, noexpansion of the endoplasmic reticulum, no swelling of mitochondria, thetight junctions normality. intestinal villus of group B were few and scatteredand disorder, expansion of the endoplasmic reticulum, mitochondria swelling,tight junctions widened.Group C compared with group B slight improvement,but there are still differences compared with group A.Conclusion: Probiotics have protective effect of TNBS-induced colitisin rats mucosal barrier and its mechanism is through up-regulation expressionof tight junction protein Claudin-1and decrease the damage of the intestinalmucosa, reducing the permeability of the intestinal mucosa, so as to achieveanti-inflammatory effect. |
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